How to Improve Germination Rates with a Reptile Heat Mat Some cannabis seeds can’t be replaced. You may have a strain or cross that’s no longer available. In the past, I’ve tried to order Why Are Your Cannabis Seeds Not Germinating? Many first-time growers and inexperienced planters often experience this problem when trying to cultivate cannabis seed. They end up languishing and Development and Standardization of Rapid and Efficient Seed Germination Protocol for Cannabis sativa Cannabis seed germination is an important process for growers and researchers alike. Many
How to Improve Germination Rates with a Reptile Heat Mat
Some cannabis seeds can’t be replaced. You may have a strain or cross that’s no longer available. In the past, I’ve tried to order cannabis seeds again, only to find the breeder disappeared. Sometimes germination problems cause irreplaceable losses. On top of that, cannabis seeds can get expensive, especially small-batch seeds or experimental seed crosses, which means if seeds don’t sprout you are down significant cash.
So far I’ve had 100% germination using this method (including several seeds that were 2 years old). Learn how to do this yourself in today’s germination tutorial.
Germination problems are some of the most frustrating problems when growing cannabis. It seems like you should be able to stick a marijuana seed in some soil and let nature take care of germination. Why is it so hard to germinate seeds? What can a beginner or experienced grower do to improve germination rates? Bonus: Why are some seeds harder to germinate than others?
What cannabis seeds want during germination:
- Moisture – never let seeds dry out after they get wet, but also don’t let seeds sit in liquid water or seedlings may “drown” after they crack open the seed
- Warm – 80°F (26°C) is perfect, though any temperature between 75-85°F (24-30°C) is good
- Dark – too much light can hurt germination so it’s recommended to germinate seeds in the dark
- No touching – try not to touch or move seeds after they start germinating until they sprout
- Clean – bacteria and microscopic fungi can slow or stop germination so make sure to wash your hands before handling seeds and germinate seeds in a clean place
So what’s the best way to give this perfect germination environment? Today I want to share the cannabis germination method I use. I’ve found most germination methods work, but one of the key things that seem to help germination go faster (and reduce the number of seeds that don’t germinate) is warmth. Seeds want it to be warm but not hot, like they’re just under the surface of the soil on a sunny spring day after a light rain.
Seeds want to “feel” like they’re just under the surface in moist dirt on a warm day. Your job is to give them a better version of nature.
Over the years I’ve struggled to achieve the perfect germination temperature. In my experience, “seedling heat mats” don’t keep a consistent temperature, and they often get too hot after a few days. I’ve germinated seeds on a seedling heat mat and I’m pretty sure they got cooked, which caused them to take much longer to germinate. So how can you achieve the perfect 80°F / 26°C temperature that seeds love? Reptile heat mat!
Why a reptile heat mat?
When you keep a cold-blooded animal like a reptile as a pet, you need to provide heat to keep them warm. However, you want to make sure their home is always the perfect temperature. If it gets too cold or hot, they can die. It’s up to you as the owner to make sure they get just-the-right temperature so they’re happy and healthy.
Pet snakes and other reptiles can’t control their own temperature so they need you to provide just the right amount of warmth
That’s why reptile heat mats were first created. It’s essentially a seedling heat mat, except it has a probe with a sensor to allow you to choose the exact temperature that you want. For example, you can set the heat mat to keep the temperature a steady 80°F / 26°C and the controller will turn the heat mat on and off so it stays exactly that temperature day and night. I’ve found that using one of these mats causes seeds to germinate much faster than if they weren’t heated, but (unlike with regular seedling heat mats) the sensor prevents the mat from ever getting too hot and cooking the seeds.
This reptile heat mat took my marijuana seed germination rates from 90-95% to almost 100% germination every time.
A reptile heat mat comes with a temperature controller to automatically let you keep your seeds at the perfect temperature so seeds germinate as quickly as possible (with no chance of overheating like with a typical seedling heat mat).
What you need
- Reptile heat mat – they come in different sizes (bigger ones can produce more heat), but the 6″x8″ size mat (uses 8 watts) should be more than enough to germinate a few seeds
- Paper towels – the cheaper the better (cheap rough paper towels work better for germinating seeds than the more expensive or cloth-like paper towels)
- Kitchen plates – you’ll use these to lock in moisture and keep seeds in the dark during germination
- Seeds – Here’s where to find seeds
- Seedling plugs (optional) – Seedling plugs like Rapid Rooters are a great place to put your newly sprouted seeds before planting them in their final home. Or just put germinated seeds directly into the soil.
Wet paper towel method – germinate seeds between wet paper towels
Use another plate on top to lock in moisture and then set up the heat mat underneath to maintain the perfect temperature (full germination tutorial below)
100% germination almost every time!
1.) Choose seeds
When starting with feminized seeds (which means all plants will be female and make buds) I germinate at least one extra seed per strain just in case a seed doesn’t germinate. If I’m sprouting non-feminized (“regular”) seeds, I germinate 4-5 seeds per strain to ensure there’s at least one female plant. Even though about half the plants from regular seeds should be female, sometimes you get unlucky and get a lot of male plants (which don’t make buds). If you don’t want to worry about male or female plants, I recommend sticking to feminized seeds so every plant ends up being female.
2.) Set up paper towels and plates
Cut your paper towels to fit your plates (if any paper towel is sticking out, it will dry everything out) and label them with the names of seeds.
Prepare paper towels, plates, and label with your chosen strains
2.) Set up reptile heating mat
In order to get everything working properly, you need to find a way to get the probe near the seeds, but not in with them or it will let light in. I’ve found the best way to do this is to place down the mat, and use a third plate upside down so you have a great place to put the probe, then set your germinating seeds in their plate on top. So you will have 3 plates in total. One to attach the probe to, and two more to encase your seeds.
Set down heating matt, and attach probe to plate.
Set the plate down. At this point, the probe is just above the heating mat. Perfect!
Add your prepared plate on top. Now it’s got the heating mat underneath, yet still an extra layer of protection to cause all the heat to radiate equally
3.) Place seeds next to their labels
Be careful not to let them roll around or you may lose track of which is which
Add seeds to their proper place. Be extra careful not to let them roll around or you won’t know which one is which after they’ve germinated. Here’s an example.
4.) Add water and cover
Gently add a little clean water, making sure to get the paper towels wet without accidentally moving the seeds around. Then take a single sheet of paper towel and add it on top. It should quickly become wet too. You want everything to seem wet, but there shouldn’t be extra water sloshing around. It shouldn’t appear too shiny. If there is visible water (for example you see water moving if you gently tip the plate), carefully dab it off with another paper towel. You want your environment totally moist but still solid.
Add water (gently without letting seeds roll around) then put a single extra sheet of paper towel on top. If you only use a single sheet of paper on top, you will still be able to see what’s underneath pretty well, as pictured here. This lets you see when seeds are germinated without disturbing them.
Having just one sheet of paper will ensure seeds get air and let you see whether they’ve germinated without having to disturb them. Seal the moisture in with another plate. Make sure no edges of the paper towels are sticking out around the edges or they will slowly wick out all the water and possibly dry out your seeds.
Seal the moisture in with another plate. Make sure no paper towels stick out the side or they’ll wick away moisture and dry everything out.
5.) Set the reptile heater to 83°F / 28°C
This isn’t an exact number (anything around 80-85° / 26-30°C does awesome), but it’s the temperature I choose. I figure a little heat is lost through the plate and it likely gets the seeds at a nice 80°F.
This is what the controller looks like
I set the reptile heater to 83°F / 28°C, but anything around that range works great. Here’s a quick video showing how to set the controller. Hold the “set” button until it blinks, set the temperature you want, then hit “set” again. You’re good to go!
If you check back in about an hour, the water should feel tepid or just slightly warm (it will never feel hot). If the water feels cold, then turn up the reptile heater a few degrees as you may be losing heat from your specific plate or your air is just a bit colder than mine.
Check on the seeds once after an hour and touch the paper towel in the middle to make sure it feels lukewarm (not hot or cold). If so, you’re good! Close it back up and try not to peak again for at least 24 hours. I know it’s hard not to check on them every hour (at least it is for me), but they will germinate better if they’re undisturbed in the dark.
6.) Seeds usually sprout in 1-3 days
Try not to check on them for 1-2 days except to make sure the temperature is right and no paper towel is sticking out the sides. Seeds like to be left alone during germination.
Before sprouting, this is what the seeds look like under the paper towel (little black dots). If you see this, cover them again and check back tomorrow.
Since you only used a single sheet of paper towel on top, you should be able to see if at least some of the seeds have germinated without having to disturb them. If you don’t see roots yet, just close the plate up and check once a day from now on. Some seeds, especially older ones (or certain strains) can take several days to germinate.
This is how seeds look once they have sprouted (through the single sheet of paper towel on top). You can faintly see their roots, and sometimes you can ever see yellow leaves. Note: the leaves are fully formed (but yellow) in the seed and break free during the germination process.
And here’s what the seedlings look like after gently peeling off the top layer
It’s normal if new seedlings look yellow (the leaves are always yellow inside the shell, but turn green once they get exposed to light)
7.) Put germinated seeds in their next home
At this point, you can put your seeds directly into their next home. I like to put seeds in Rapid Rooters because that gives me a few more days to examine all the seedlings and see which ones are growing the best before picking the winners for their final home.
You can also put germinated seeds directly into soil or coco, just be gentle and try to plant seeds with their roots down.
Put your sprouted seedling inside, with the seed head just under the surface
Gently close the Rapid Rooter around the seedling and put it in a tray or shot glass (or any way to hold it upright while the root is growing). If the seed/seedling seems loose like it might move around, pick a little piece off a Rapid Rooter and gently put it in the hole to make sure the seed head is totally secure (you don’t want it moving around while the root is still growing into the seedling plug).
Put Rapid Rooters under gentle light in a warm place (for example in your regular grow spot with the light 2-3x further than normal, or near a sunny window)
Soon your sprouts will be seedlings
Once seed leaves are about the width of the Rapid Rooter, they’re ready to be planted in their final home
Why Are Your Cannabis Seeds Not Germinating?
Many first-time growers and inexperienced planters often experience this problem when trying to cultivate cannabis seed. They end up languishing and wondering why their effort is not yielding since a cannabis seed has a pretty high germination rate (99%). However, many things might not promote the germination process.
Generally, cannabis seeds are powerful and grow pretty fast. If the seeds do not germinate, there is likely a problem with the germination method. This article will explore many things that can go wrong when trying to cultivate cannabis seeds.
The chances of having a viable seed increase significantly if the genetics is good. This is pretty important as it can avoid wasting energy and effort trying to germinate a bad seed. When you buy good quality seeds from trusted banks, you can breathe a sigh of relief, knowing that your actions will not be wasted.
As a result, it is not a good idea to grow seeds that you stumbled on inside a cannabis bag you got. Such seeds could be infertile and may not germinate. If it eventually germinates, you might not like cannabis. Make sure to buy the right seed from reputable stores and increase the chances of germination.
Direct Soil Germination
This is one of the top reasons your cannabis seed is not germinating. If you do not water the substrate before sowing the seeds and water afterward, there is a high probability that the seeds will not grow. This is not surprising as the seeds could be buried too deep, and adding water after sowing worsens this.
You can obtain good results by germinating the seeds in jiffy pellets, peat plugs, or using kitchen paper and later transplant them to a pot or the soil when the small seedlings come out. Make sure to care for your tender plant at this stage. Limit irrigation, support the right temperature and use Northern lights when growing indoors.
Wrong Temperature and Humidity
The ideal temperature for germinating cannabis seeds is high humidity and high-temperature levels. It is essential for people in tropical countries to provide external support of heat to get the temperature within reasonable and acceptable ranges. This is where heated greenhouses come in. Not only do they provide the right temperature, but they can get the humidity level to the ideal range essential for germinations. A cannabis seed needs 70% relative humidity and an average temperature of about 270C for germination.
If the temperature and relative humidity ranges are lower than the ideal value specified, the growth rate will be slow and unsuccessful. Excessive range, on the other hand, can lead to fungal problems.
Seeds Planted too Deep.
The amount of energy available to a germinating cannabis seed is limited. As a result, if planted too deep, it might not have the required energy to make its way up. A seed planted underground will have no light and is not producing food yet since no photosynthesis. Planting seed too deep creates extra stress for the seedling as migration to the surface becomes an issue.
As a result, you should plant your cannabis seed around 2 to 3 cm down or half inches deep. With this, they have enough room to grow taproots. This might be one of the reasons your cannabis seed didn’t grow.
Using Old or Unsterilized Pots and Soils
A fungus is one of the greatest enemies that might affect the germination of cannabis seed. If you reuse soil and it is not sterilized, there is a high chance of mold and other unhealthy organisms like insects and bacteria. As a result, the seeds may not sprout at all. Even if they grow, these organisms might kill them after a couple of days. For instance, a sprouting seedling might suddenly bend and change color to brown in a process known as damping off. This is common when you supply the seed with too much water when there is poor drainage or the aeration is poor.
Due to this, consider planting seedlings in a sterilized pot as the tendency of containing harmful organisms is low. Also, make sure the containers are clean as it can reduce the tendency of fungus.
Planting lots of Seed in one Pot
Planting a lot of seed in a single container might seem like a good idea due to the ability to save cost. This is like shooting yourself in the leg, as it might lead to a wasted effort. Even if the seeds germinate, they will compete for limited space, nutrients, and water. A cannabis plant needs enough space for its root to travel down without restriction to prevent severe intermodal distance.
Limited space translates to severe competition not only for nutrients but light as well. This is not a good idea if you want to get the best from your cannabis seed. If the seed eventually germinates, they will be weak, which will affect the plants and overall output.
Cannabis Seed left underwater for a long time.
Some growers prefer to soak their cannabis seeds before planting them. This is an essential step to break the dormancy and soften the shell to prepare the seed for planting. With a softshell, the taproot will easily break the body and push its way out of the shell. Typically, the seed will be put inside a glass of water and left until it sinks before the grower plants it. However, this is not as simple as it sounds.
A cannabis seed needs to be continually breathing, and a fully submerged seed will not get the necessary air supply. This might kill the seed if left for long inside the water. Once the seed sinks, take them inside the paper towel as soon as possible.
There are many intricacies involved in germinating a cannabis seed. Getting it wrong with any of the systems might not make the cannabis seed grow. You can arm yourself with these essential tips to ensure that your cannabis plants germinate. Use these tips as a guide when trying to germinate your cannabis seed. Also, if your seed does not germinate, this guide can help you troubleshoot.
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Development and Standardization of Rapid and Efficient Seed Germination Protocol for Cannabis sativa
Cannabis seed germination is an important process for growers and researchers alike. Many biotechnological applications require a reliable sterile method for seed germination. This protocol outlines a seed germination procedure for Cannabis sativa using a hydrogen peroxide (H2O2) solution as liquid germination media. In this protocol, all three steps including seed sterilization, germination, and seedlings development were carried out in an H2O2 solution of different concentrations; 1% H2O2 solution showed the fastest and the most efficient germination. This protocol also exhibited high germination efficiency for very old cannabis seeds with lower viability. Overall, this protocol demonstrates superior germination compared to water control and reduces the risk of contamination, making it suitable for tissue culture and other sensitive applications.
Keywords: Cannabis sativa, Rapid germination, Hydrogen peroxide, Seed sterilization, Seedling development
Cannabis sativa, otherwise known as marijuana or hemp, is an annual primarily dioecious flowering plant in which male/female sex is determined by heteromorphic chromosomes (X and Y) ( Gaudet et al., 2020 ). Cannabis is grown for a variety of agricultural uses; nearly all parts of cannabis plant are used, seeds for food, stem for fiber, and flowers/leaves for medicine. Flowers produce a mix of cannabinoids and aromatic compounds valued for their therapeutic and recreational effects ( Chandra et al., 2017 ). Cannabis plants are propagated either clonally through cuttings or via seed germination. Seed germination is very important for researchers, breeders, and growers alike, especially since seeds from elite cultivars can be very expensive and valuable. Additionally, older seeds may have a reduced germination rate while bacterial and fungal contamination can compromise germination, especially when seeds are germinated for tissue culture propagation. To address these issues, we have developed a rapid, sterile, and efficient seed germination protocol using a 1% hydrogen peroxide (H2O2) solution. In this protocol, all three steps including seed sterilization, germination, and seedlings development were carried out in a 1% H2O2 solution. This presents a significant advantage over other sterilants, such as mercuric chloride or bleach, which require additional washing of seeds and a separate germination step on MS solid medium. Our protocol resulted in faster germination and increased seed germination percentage as compared to water control, with no bacterial or fungal contamination, making it suitable for tissue culture and other sensitive applications. In comparison to previous germination methods which take between 4-7 days for radicle appearance and 5-15 days for seedling development ( Wielgus et al., 2008 and references therein), our germination method resulted in radicle appearance in 1 day and allowed us to obtain cannabis seedlings in a very short period (3-7 days) with minimal efforts. This protocol is also very efficient for germination of very old cannabis seeds with lower viability.
Materials and Reagents
All seeds were harvested in our laboratory. Blueberry seeds were not older than 6 months, when employed in the experiments. Finola and X59 seeds were more than 5 years old.
Growth chamber (Sanyo MLR-350, catalog number: 859-600-06): 24 °C, 18 h light/6 h dark cycle, light intensity 200 μmol·m -2 ·sec -1
Seed germination assay
Soak seeds overnight in various concentrations of hydrogen peroxide solution (liquid germination media or germination solutions) as well as in sterile water control (H2O, 1% H2O2, 3% H2O2, 5% H2O2, or 10% H2O2) in 15 or 50 ml screw-cap (Falcon tube). Falcon tubes with submerged seeds in various germination solutions were kept in the dark at room temperature.
Next day, record the percentage of germinated seeds in germination solution (appearance of radicle is considered as germination event) and add fresh respective germination solution after removal of old solution simply by pouring out.
Keep seeds soaked in the same solution for 3 more days in the dark at room temperature and record the percentage of germinated seeds every day.
Thereafter, germinated seeds/seedlings were transferred with or without seed coats from H2O2 solution to MS medium plates to observe the growth of H2O2 solution-germinated seeds/seedlings on MS medium. To transfer, first germinated seeds/seedlings were poured together with H2O2 solution from the Falcon tube to the empty petri plate. Then seedlings were transferred to sterile paper by using forceps to remove excess H2O2 solution. Finally, the germinated seeds/seedlings were transferred to MS media plate by using forceps. The whole transfer process has been carried out in the laminar flow hood.
Parafilm sealed MS medium plates with germinated seeds/seedlings are then transferred to the growth chamber (24 °C, 18 h light/6 h dark cycle and light intensity 200 μmol·m -2 ·sec -1 ) for 3 days to observe the growth and survival of H2O2 solution germinated seeds/seedlings on MS medium.
The H2O2 solution-germinated seeds/seedlings growth was also observed in soil. Pro-Mix HP Mycorrhizae Growing Medium used for soil experiment. The cannabis seeds were soaked in the H2O2 solution (germination solutions) for four days and thereafter, germinated seeds/seedlings were transferred from H2O2 solution to soil pot (Pro-Mix HP Mycorrhizae Growing Medium) to observe the growth and survival of H2O2 solution germinated seeds/seedlings on soil. The soil pots were transferred to the growth chamber (24 °C, 18 h light/6 h dark cycle and light intensity 200 μmol·m -2 ·sec -1 ). The photographs were taken on day 12.
Mean seed germination percentage under various concentrations of H2O2 solution as well as water control were calculated in an excel sheet. Data were shown as mean ± SE.
In this study, we have described a rapid and efficient seed germination protocol for Cannabis sativa. The brief description of this protocol has been reported in Sorokin et al. (2020) . In the current study, we have standardized the optimum concentration of hydrogen peroxide (H2O2) solution media for efficient sterilization and rapid germination. We have tested various concentrations of H2O2 solution as well as sterile water control (H2O, 1% H2O2, 3% H2O2, 5% H2O2, or 10% H2O2) for sterilization and germination efficiency. All three steps of germination (seed sterilization, germination, and seedlings development) were carried out in various concentrations of H2O2 solution and seeds were kept in liquid media for four days. Hydrogen peroxide presents several significant advantages over mercuric chloride or bleach sterilants, which require additional seed washing, and separate germination/seedling development step in Murashige and Skoog (MS) agar medium ( Sorokin et al., 2020 ). The 1% H2O2 solution showed rapid and higher germination than higher H2O2 concentrations solution and water control at day 1 ( Figure 1 ). On day 1, 1% H2O2 solution exhibited 82.5% germination as compared to 22.5% germination for 3% H2O2 group, 17.5% germination for 5% H2O2 group and 47.5% germination in water control group ( Figure 1B ). Interestingly, 10% H2O2 did not show any germination on day 1 due to its toxic effect ( Figure 1 ). In 1% H2O2 solution, radicle appearance (germination) occurred within 24 h and seedling development (two fully developed cotyledons and two immature true leaves stage) occurred in 72-96 h ( Figure 1A ). In comparison to previous germination methods which take between 4-7 days for radicle appearance and 5-15 days for seedling development ( Wielgus et al., 2008 and references therein), our germination method resulted in radicle appearance in 1 day and allowed us to obtain cannabis seedlings in a very short period (3-7 days) with minimal efforts ( Figures 1 -2). Considering the possible toxic effect of H2O2 (since germinated seeds/seedlings stayed continuously in H2O2 solution for 4 days), we have checked further survival of germinated seeds/seedlings on MS media and soil ( Figures 2 -3). On MS media, 1% H2O2 solution seedlings survived better than other treatments ( Figure 2 ). The water germinated seeds exhibited contamination and did not survive on MS media ( Figure 2 ). Similarly, due to the toxic effect of higher concentration of H2O2, the 10% H2O2 germinated seeds did not survive on MS media ( Figure 2 ). The 1% H2O2 solution seedlings also survived well on soil ( Figure 3 ). Apart from this, we have also tested our method for more than 5-years old cannabis seeds with lower viability, which demonstrated that 1% H2O2 solution medium exhibited a very high germination percentage (~50%) as compared to water control (~10%) ( Figure 4 ). In conclusion, we have developed a rapid and efficient method for C. sativa seed germination under sterile conditions for tissue culture and other sensitive applications.
Germination of 6-month-old seeds of Blueberry variety in various concentrations of hydrogen peroxide solution and water control.
A. Representative photographs of germinated seeds/seedlings in the H2O2 solution of various concentrations or water control on day 1 to day 4. B. Comparison of germination percentage between the various concentrations of H2O2 solution or water control. Data are shown as mean ± SE (n = 4). In each replicate, 30 seeds were used.
Representative photographs of growth and survival of H2O2 solutions germinated seeds/seedlings of Blueberry variety on MS media.
The Blueberry variety seeds were soaked in the H2O2 solution (germination solutions) for four days and thereafter, germinated seeds/seedlings were transferred from H2O2 solution to MS medium plates to observe the growth and survival of H2O2 solution germinated seeds/seedlings on MS medium. The photographs were taken at day 0 (just after transfer to MS medium plates), day 1 (after 24 h of the transfer to MS medium plates), and day 3 (after 72 h of the transfer to MS medium plates) on MS media.
Representative photograph of Blueberry variety young plantlet growing in soil (Pro-Mix HP Mycorrhizae Growing Medium).
The Blueberry variety seeds were soaked in the H2O2 solution (germination solutions) for four days and thereafter, germinated seeds/seedlings were transferred from H2O2 solution to soil pot (Pro-Mix HP Mycorrhizae Growing Medium) to observe the growth and survival of H2O2 solution germinated seeds/seedlings on soil. The photographs were taken on day 12.
Germination of 5-years old seeds of Finola and X59 varieties in 1% hydrogen peroxide solution and water control.
Comparison of germination percentage between 1% H2O2 solution media and water control. Data are shown as mean ± SE (n = 5). In each replicate, around 30 seeds were used.
4.43 g Murashige & Skoog Basal Medium with Vitamins
Adjust pH to 5.7 with KOH and sterilize by autoclaving at 121 °C for 40 min. 25 ml of MS media on each Petri plate.
This protocol is derived from Sorokin et al. (2020). We thank the Natural Sciences and Engineering Research Council of Canada (NSERC) and MITACS for funding our work.
The authors declare that they have no competing interests.
Readers should cite both the Bio-protocol article and the original research article where this protocol was used.
1. Chandra S., Lata H. and ElSohly M. A.(2017). Cannabis sativa L.-botany and biotechnology. Chandra, S., Lata, H. and ElSohly, M. A.(Eds.). Springer International Publishing: Cham, Switzerland. ISBN: 9783319545639. [Google Scholar]
2. Gaudet D., Yadav N. S., Sorokin A., Bilichak A. and Kovalchuk I.(2020). Development and optimization of a germination assay and long-term storage for Cannabis sativa pollen . Plants 9 : 665. [PMC free article] [PubMed] [Google Scholar]
3. Sorokin A., Yadav N. S., Gaudet D. and Kovalchuk I.(2020). Transient expression of the β-glucuronidase gene in Cannabis sativa varieties . Plant Signal Behav 15 ( 8 ): 1780037. [PMC free article] [PubMed] [Google Scholar]
4. Wielgus K., Luwanska A., Lassocinski W. and Kaczmarek Z.(2008). Estimation of Cannabis sativa L. tissue culture conditions essential for callus induction and plant regeneration . J Nat Fibers 5 : 199-207. [Google Scholar]